Abstract 4151: Creatine and GAMT promote tumor stemness and malignancy and can be used as biomarkers for colorectal cancer recurrence

Abstract BACKGROUND: The role of creatine, a common nutritional supplement, in tumorigenesis and development has gained increasing attention. Previous studies have shown that it has a dual effect on tumorigenesis and recent studies suggested that it promotes colorectal cancer (CRC) metastasis. However, the role creatine metabolism plays in tumor malignancy and recurrence is still unclear and identifying the mechanism could provide new strategies for the treatment of CRC. METHODS: Targeted metabolomics analysis of creatine was dynamically performed on plasma samples of 160 CRC patients including 30 relapse cases. GAMT, the key enzyme in creatine synthesis, was investigated in TCGA and ICGC-ARGO database (a large multi-omics study of CRC in China conducted by the sixth affiliated hospital of Sun Yat-sen University). The effect of creatine and GAMT on CRC stemness and its mechanism were investigated in CRC cell lines, patient-derived organoids, and patient-derived xenografts. Limited proteolysis-mass spectrometry (LiP-MS) and RNA-sequencing was performed to illustrate creatine-protein interaction and the regulation of relative signal pathways. RESULTS: In recurrent CRC cases, creatine level gradually increased over time after the first radical surgery until recurrence. Meanwhile, GAMT expression was found significantly higher in patients who were going to relapse, both in TCGA and ICGC-ARGO database. Patients with higher GAMT expression had worse prognosis. The addition of creatine inhibitor can effectively restrain CRC activity and proliferation both in vitro and in vivo. GAMT knockdown significantly reduced the self-renewal ability of CRC cells, and tumor stemness pathways were up-regulated, which can be reversed by adding creatine to the culture. CONCLUSIONS: Creatine is associated with CRC recurrence through promoting tumor stemness and malignancy. Therapeutic strategies targeting creatine metabolism have the potential to reduce CRC recurrence. Citation Format: Zhengran Zhou, Peishan Hu, Zerong Cai, Yufeng Chen, Xiaojian Wu, Xuanhui Liu. Creatine and GAMT promote tumor stemness and malignancy and can be used as biomarkers for colorectal cancer recurrence [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 4151.